Specific interference shRNA-expressing plasmids inhibit Hantaan virus infection in vitro and in vivo

Liu, Yuan-yuan; Chen, Liang-jun; Zhong, Yan; Shen, Meng-xin; Ma, Nian; Liu, Bing-yu; Luo, Fan; Hou, Wei; Yang, Zhan-qiu; Xiong, Hai-rong

The RNAi pSilencer-S and pSilencer-M plasmids were constructed, and their antiviral effects were further evaluated by detecting the viral protein synthesis and RNA transcript and progeny virus titers in the HTNV-infected cells. The transfection efficiencies were evaluated according to the GFP expression of the co-transfected plasmid pEGFP-C1 and reached approximately 70%-80%. Treatment with pSilencer-S or pSilencer-M at concentrations of 1, 2, and 4 μg significantly decreased viral the antigen expression in the HTNV-infected Vero-E6 cells at 3 dpi ( Figure 2 ). The percentages of positive cells declined with increases in the dose of transfected RNAi plasmids ( Figure 2H and 2I) . Only 14.69%±1.78% of the cells were positive for nucleocapsid protein staining in the 2-μg pSilencer-S treatment group compared to 56.8%±3.21% in the pSilencer-blank treatment group ( Figure 2H) . Similarly, the corresponding glycoprotein G2 positive staining rates were 14.8%±2.41% and 58.33%±6.24% in the 2-μg pSilencer-M treatment group ( Figure 2I ). These results indicate that the 2-μg plasmid transfection achieved the most effective inhibition of viral protein synthesis.